Structural Characterization and Dynamics of AdhE Ultrastructures from Clostridium Thermocellum: A Containment Strategy for Toxic Intermediates

Ziegler, Samantha J.; Knott, Brandon C.; Gruber, Josephine N.; Hengge, Neal N.; Xu, Qi; Olson, Daniel G.; Romero, Eduardo E.; Joubert, Lydia M.; Bomble, Yannick J.

doi:10.7554/eLife.96966.2

Structural Characterization and Dynamics of AdhE Ultrastructures from Clostridium Thermocellum: A Containment Strategy for Toxic Intermediates

Journal Article · Thu May 15 04:00:00 EDT 2025

DOI:https://doi.org/10.7554/eLife.96966.2· OSTI ID:2568050

Ziegler, Samantha J.; Knott, Brandon C.; Gruber, Josephine N.; Hengge, Neal N.; Xu, Qi; Olson, Daniel G.; Romero, Eduardo E.; Joubert, Lydia M.; Bomble, Yannick J.

Clostridium thermocellum, a cellulolytic thermophilic anaerobe, is considered by many to be a prime candidate for the realization of consolidated bioprocessing (CBP) and is known as an industry standard for biofuel production. C. thermocellum is among the best biomass degraders identified to date in nature and produces ethanol as one of its main products. Many studies have helped increase ethanol titers in this microbe, however ethanol production using C. thermocellum is still not economically viable. Therefore, a better understanding of its ethanol synthesis pathway is required. The main pathway for ethanol production in C. thermocellum involves the bifunctional aldehyde-alcohol dehydrogenase (AdhE). To better understand the function of the C. thermocellum AdhE, we used cryo-electron microscopy (cryo-EM) to obtain a 3.28 A structure of the AdhE complex. This high-resolution structure, in combination with molecular dynamics simulations, provides insight into the substrate channeling of the toxic intermediate acetaldehyde, indicates the potential role of C. thermocellum AdhE to regulate activity and cofactor pools, and establishes a basis for future mutagenesis studies.

Research Organization:: National Renewable Energy Laboratory (NREL), Golden, CO (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

DOE Contract Number:: AC36-08GO28308; AC36-08GO28308

OSTI ID:: 2568050

Report Number(s):: NREL/JA-2700-87838; MainId:88613; UUID:4e9fa7c4-acfe-4388-ad2d-2352bfa846a8; MainAdminId:73330

Country of Publication:: United States

Language:: English

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